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Roomans, Godfried M.
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Publications (10 of 18) Show all publications
Hussain, R., Shahror, R., Karpati, F. & Roomans, G. M. (2015). Glucocorticoids can affect Pseudomonas aeruginosa (ATCC 27853) internalization and intracellular calcium concentration in cystic fibrosis bronchial epithelial cells. Experimental Lung Research, 41(7), 383-392
Open this publication in new window or tab >>Glucocorticoids can affect Pseudomonas aeruginosa (ATCC 27853) internalization and intracellular calcium concentration in cystic fibrosis bronchial epithelial cells
2015 (English)In: Experimental Lung Research, ISSN 0190-2148, E-ISSN 1521-0499, Vol. 41, no 7, p. 383-392Article in journal (Refereed) Published
Abstract [en]

Background and Objective: Glucocorticoids (GCs) are anti-inflammatory agents, but their use in cystic fibrosis (CF) is controversial. In CF, the early colonization with Pseudomonas aeruginosa is mainly due to nonmucoid strains that can internalize, and induce apoptosis in the epithelial cells. Uptake of P. aeruginosa by the epithelial cells and subsequent apoptosis may prevent colonization of P. aeruginosa in CF airways. In the airway epithelia, several other biological effects, including an anti-secretory role by decreasing intracellular Ca2+ concentration have been described for this anti-inflammatory drug. However, the effects of GCs on the nonmucoid P. aeruginosa internalization and intracellular Ca2+ in CF bronchial epithelial cells have not been evaluated.

Methods: We used cultured human CF bronchial airway epithelial cell (CFBE) monolayers to determine P. aeruginosa internalization, apoptosis, and intracellular Ca(2+)concentration in CF bronchial epithelial cells. Cells were treated with IL-6, IL-8, dexamethasone, betamethasone, or budesonide.

Results: GCs in co-treatments with IL-6 reversed the effect of IL-6 by decreasing the internalization of P. aeruginosa in the CFBE cells. GCs decreased the extent of apoptosis in CFBE cells infected with internalized P. aeruginosa, and increased the intracellular Ca2+ concentration.

Conclusion: These findings suggest that if internalization of P. aeruginosa reduces infection, GC therapy would increase the risk of pulmonary infection by decreasing the internalization of P. aeruginosa in CF cells, but GCs may improve airway hydration by increasing the intracellular Ca2+ concentration. Whether the benefits of GC treatment outweigh the negative effects is questionable, and further clinical studies need to be carried out.

Place, publisher, year, edition, pages
Taylor & Francis, 2015
Keywords
Cystic fibrosis, glucocorticoids, internalization, intracellular Ca2+, Pseudomonas aeruginosa
National Category
Respiratory Medicine and Allergy
Research subject
Pulmonary Medicine
Identifiers
urn:nbn:se:oru:diva-48764 (URN)10.3109/01902148.2015.1046199 (DOI)000369738800003 ()26151838 (PubMedID)2-s2.0-84942123541 (Scopus ID)
Funder
Swedish Heart Lung Foundation
Available from: 2016-02-29 Created: 2016-02-29 Last updated: 2017-11-30Bibliographically approved
Hussain, S., Varelogianni, G., Särndahl, E. & Roomans, G. M. (2015). N-acetylcysteine and azithromycin affect the innate immune response in cystic fibrosis bronchial epithelial cells in vitro. Experimental Lung Research, 41(5), 251-260
Open this publication in new window or tab >>N-acetylcysteine and azithromycin affect the innate immune response in cystic fibrosis bronchial epithelial cells in vitro
2015 (English)In: Experimental Lung Research, ISSN 0190-2148, E-ISSN 1521-0499, Vol. 41, no 5, p. 251-260Article in journal (Refereed) Published
Abstract [en]

Background and objective: We have previously reported that N-acetylcysteine (NAC), ambroxol and azithromycin (AZM) (partially) correct the chloride efflux dysfunction in cystic fibrosis bronchial epithelial (CFBE) cells with the ΔF508 homozygous mutation in vitro.

Methods: In the present paper, we further investigated possible immunomodulatory effects of these drugs on the regulation of the innate immune system by studying the expression of the cytosolic NOD-like receptors NLRC1 and NLRC2, and interleukin (IL)-6 production in CFBE cells.

Results: Under basal conditions, PCR and Western Blot data indicate that the NLRC2 receptor has a reduced expression in CF cells as compared to non-CF (16HBE) cells, but that the NLRC1 expression is the same in both cell lines. AZM significantly upregulated NLRC1 and NLRC2 while NAC upregulated only NLRC2 receptor expression in CF cells. Reduced basal IL-6 production was found in CF cells as compared to non-CF cells. MDP (an NLRC2 agonist), NAC and AZM, but not Tri-DAP (an NLRC1 agonist), increased IL-6 production in CF cells, indicating that in CF cells IL-6 upregulation is independent of NLRC1, but involves the activation of NLRC2.

Conclusion: Overall, the results indicate that NAC and AZM not only can correct the chloride efflux dysfunction but also have a weakly strengthening effect on the innate immune system.

Place, publisher, year, edition, pages
London, UK: Informa Healthcare, 2015
Keywords
Chloride efflux activators; Cystic fibrosis; NOD-like receptors
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Respiratory Medicine and Allergy
Research subject
Pulmonary Medicine
Identifiers
urn:nbn:se:oru:diva-41292 (URN)10.3109/01902148.2014.934411 (DOI)000357350400002 ()25058850 (PubMedID)2-s2.0-84930853087 (Scopus ID)
Funder
Swedish Heart Lung Foundation
Note

Funding Agencies:

Swedish Cystic Fibrosis Association

Nyckelfonden-the Foundation for Medical Research at Örebro University Hospital

Available from: 2015-01-14 Created: 2015-01-14 Last updated: 2018-06-26Bibliographically approved
Hussain, R., Hugosson, S. & Roomans, G. M. (2014). Isolation and culture of primary human nasal epithelial cells from anesthetized nasal epithelia. Acta Oto-Laryngologica, 134(3), 296-299
Open this publication in new window or tab >>Isolation and culture of primary human nasal epithelial cells from anesthetized nasal epithelia
2014 (English)In: Acta Oto-Laryngologica, ISSN 0001-6489, E-ISSN 1651-2251, Vol. 134, no 3, p. 296-299Article in journal (Refereed) Published
Abstract [en]

Conclusion: Using a local anesthetic agent before obtaining nasal biopsies by nasal brushing makes the sampling procedure smooth, avoids lacrimation, nasal itching/irritation, and/or sneezing and provides enough viable cells to establish primary cultures.

Objectives: To examine the use of local anesthesia to avoid the irritation experienced by the subject when nasal biopsies are obtained by nasal brushing in order to culture viable nasal epithelial cells.

Methods: Nasal epithelial cells were collected from the mid-part of the inferior turbinate of healthy volunteers by brushing with interdental brushes, after spraying a topical anesthetic on the nasal mucosa. Immunocytochemistry was performed to assess the purity of epithelial cells.

Results: Cell samples ranging from 1.16 x 10(5) to 3.06 x 10(5) cells/per sample were obtained. Of 11 samples, 7 formed confluent cultures, while the remaining 4 samples showed only patches of epithelial cells. Neither fungal nor bacterial contamination posed a problem. Immunocytochemistry of the cytospin slides confirmed the presence of epithelial cells in the cultures. No adverse effects were experienced by the volunteers.

Place, publisher, year, edition, pages
London, United Kingdom: Informa Healthcare, 2014
Keywords
Nasal brush biopsy, airway epithelia, lidocaine
National Category
Medical and Health Sciences Otorhinolaryngology
Research subject
Medicine
Identifiers
urn:nbn:se:oru:diva-34502 (URN)10.3109/00016489.2013.859396 (DOI)000331829700014 ()24359095 (PubMedID)2-s2.0-84894384458 (Scopus ID)
Funder
Swedish Heart Lung FoundationSwedish Research Council
Available from: 2014-03-31 Created: 2014-03-31 Last updated: 2018-06-05Bibliographically approved
Roomans, G. M. (2014). Pharmacological treatment of the basic defect in cystic fibrosis. Cell Biology International, 38(11), 1244-1246
Open this publication in new window or tab >>Pharmacological treatment of the basic defect in cystic fibrosis
2014 (English)In: Cell Biology International, ISSN 1065-6995, E-ISSN 1095-8355, Vol. 38, no 11, p. 1244-1246Article, review/survey (Refereed) Published
Abstract [en]

Cystic fibrosis (CF) is a genetic disease due to a mutation in the cystic fibrosis transmembrane conductance regulator (CFTR), a chloride channel in epithelial cells. There are about 1900 mutations, divided in several groups, for example, stop mutations, mutations affecting the permeability of the channel, and mutations in which the mutated CFTR is recognized as abnormal and destroyed. Pharmacological treatment has become possible for stop mutations (about 10% of the patients), and for a rare mutation affecting channel permeability. For the majority of patients, however, that have a mutation in which the mutated CFTR is destroyed on its way to the cell membrane, research is still in progress, although a number of compounds have been identified that (at least partly) corrects the error in chloride transport.

Place, publisher, year, edition, pages
West Sussex, United Kingdom: John Wiley & Sons, 2014
Keywords
Ion channels, membrane transport, pharmacology, respiration
National Category
Cell and Molecular Biology
Research subject
Cell Research
Identifiers
urn:nbn:se:oru:diva-38514 (URN)10.1002/cbin.10312 (DOI)000342798200002 ()24809326 (PubMedID)2-s2.0-84908221675 (Scopus ID)
Available from: 2014-11-11 Created: 2014-11-11 Last updated: 2018-01-11Bibliographically approved
Roomans, G. M. & Dragomir, A. (2014). X-Ray Microanalysis in the Scanning Electron Microscope (3ed.). In: John Kuo (Ed.), Electron microscopy: methods and protocols (pp. 639-661). New York, USA: Humana Press
Open this publication in new window or tab >>X-Ray Microanalysis in the Scanning Electron Microscope
2014 (English)In: Electron microscopy: methods and protocols / [ed] John Kuo, New York, USA: Humana Press, 2014, 3, p. 639-661Chapter in book (Refereed)
Abstract [en]

X-ray microanalysis conducted using the scanning electron microscope is a technique that allows the determination of chemical elements in bulk or semi-thick specimens. The lowest concentration of an element that can be detected is in the order of a few mmol/kg or a few hundred parts per million, and the smallest amount is in the order of 10(-18) g. The spatial resolution of the analysis depends on the thickness of the specimen. For biological specimen analysis, care must be taken to prevent displacement/loss of the element of interest (usually ions). Protocols are presented for the processing of frozen-hydrated and freeze-dried specimens, as well as for the analysis of small volumes of fluid, cell cultures, and other specimens. Aspects of qualitative and quantitative analysis are covered, including limitations of the technique.

Place, publisher, year, edition, pages
New York, USA: Humana Press, 2014 Edition: 3
Series
Methods in molecular biology, ISSN 1064-3745 ; 1117
Keywords
X-ray microanalysis, Scanning electron microscopy, Peak-to-background ratio, Frozen-hydrated specimen, Sephadex beads, Cell culture, Airway surface liquid
National Category
Biochemistry and Molecular Biology
Research subject
Biochemistry; Molecular Biology
Identifiers
urn:nbn:se:oru:diva-37564 (URN)10.1007/978-1-62703-776-1_28 (DOI)24357383 (PubMedID)2-s2.0-84934434731 (Scopus ID)978-1-62703-776-1 (ISBN)978-1-62703-775-4 (ISBN)
Available from: 2014-10-07 Created: 2014-10-07 Last updated: 2017-10-17Bibliographically approved
Hussain, R., Oliynyk, I., Roomans, G. M. & Björkqvist, M. (2013). Modulation of ENaC, CFTR, and iNOS expression in bronchial epithelial cells after stimulation with Staphylococcus epidermidis (94B080) and Staphylococcus aureus (90B083). Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), 121(9), 814-826
Open this publication in new window or tab >>Modulation of ENaC, CFTR, and iNOS expression in bronchial epithelial cells after stimulation with Staphylococcus epidermidis (94B080) and Staphylococcus aureus (90B083)
2013 (English)In: Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS), ISSN 0903-4641, E-ISSN 1600-0463, Vol. 121, no 9, p. 814-826Article in journal (Refereed) Published
Abstract [en]

Bacteria affect the respiratory epithelium, which is covered by airway surface liquid (ASL) and mucus. Ion concentrations in the ASL are determined by the cystic fibrosis transmembrane conductance regulator (CFTR) and the epithelial Na+ channel (ENaC). Neonatal sepsis is a major risk factor for subsequent pulmonary disease in preterm newborns. Predominating are coagulase-negative staphylococci (e.g., Staphylococccus epidermidis and Staphylococccus aureus). The aim of this study was to investigate modulation of CFTR, ENaC, mucins, proinflammatory cytokines, and inducible nitric oxide synthase (iNOS) in respiratory epithelial cells after S. epidermidis 94B080 and S. aureus 90B083 exposure. Bronchial epithelial cells were incubated with S. epidermidis 94B080 and S. aureus 90B083 (neonatal blood isolates) for 1-36h. Expression of CFTR, ENaC, iNOS, and mucins was analyzed by real-time PCR and Western blotting. Release of cytokines was analyzed by ELISA, and production of NO by the Griess assay. Expression of CFTR significantly decreased after 36h incubation with S. epidermidis and more prominently with S. aureus, whereas S. epidermidis caused a significant increase in the expression of - and -ENaC. Expression of iNOS increased, but NO was not detected. Both staphylococci caused a decrease in the intracellular Ca2+ concentration. S. aureus induced increased secretion of IL-6, IL-8, and transforming nuclear factor (TNF)- in a time-dependent manner as compared with S. epidermidis. In conclusion, expression of ENaC, CFTR, and iNOS is modulated by exposure to S. aureus 90B083 and S. epidermidis 94B080. S. aureus is more potent in causing release of IL-6, IL-8, and TNF- by bronchial epithelial cells as compared with S. epidermidis. The mRNA expression for the mucus proteins MUC2, MUC5AC, and MUC5B could not be measured, neither in the presence nor in the absence of bacteria.

Place, publisher, year, edition, pages
Hoboken, USA: Wiley-Blackwell, 2013
Keywords
Airway epithelium, Staphylococcus aureus 90B083, Staphylococcus epidermidis 94B080, CFTR, ENaC, iNOS, cytokines
National Category
Medical and Health Sciences Immunology
Research subject
Medicine
Identifiers
urn:nbn:se:oru:diva-30514 (URN)10.1111/apm.12138 (DOI)000322757000003 ()23879620 (PubMedID)2-s2.0-84881558007 (Scopus ID)
Note

Funding Agencies:

Nyckelfonden of Örebro University Hospital 

Swedish Science Research Council 

Swedish Heart Lung Foundation 

Available from: 2013-08-30 Created: 2013-08-30 Last updated: 2018-05-21Bibliographically approved
Varelogianni, G., Hussain, R., Strid, H., Oliynyk, I., Roomans, G. M. & Johannesson, M. (2013). The effect of ambroxol on chloride transport, CFTR and ENaC in cysticfibrosis airway epithelial cells. Cell Biology International, 37(11), 1149-1156
Open this publication in new window or tab >>The effect of ambroxol on chloride transport, CFTR and ENaC in cysticfibrosis airway epithelial cells
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2013 (English)In: Cell Biology International, ISSN 1065-6995, E-ISSN 1095-8355, Vol. 37, no 11, p. 1149-1156Article in journal (Refereed) Published
Abstract [en]

Ambroxol, a mucokinetic anti-inflammatory drug, has been used for treatment of cystic fibrosis (CF). The respiratoryepitheliumis covered by the airway surface liquid (ASL), the thickness and composition of which is determined by Cl efflux viathe cystic fibrosis transmembrane conductance regulator (CFTR) and Naþ influx via the epithelial Naþ channel (ENaC). In cellsexpressing wt-CFTR, ambroxol increased the Cl- conductance, but not the bicarbonate conductance of the CFTR channels.Weinvestigated whether treatment with ambroxol enhances chloride transport and/or CFTR and ENaC expression in CF airwayepithelial cells (CFBE) cells. CFBE cells were treated with 100 mM ambroxol for 2, 4 or 8 h. mRNA expression for CFTR andENaC subunits was analysed by real-time polymerase chain reaction (RT-PCR); protein expression was measured by Westernblot. The effect of ambroxol on Cl− transport was measured by Cl− efflux measurements with a fluorescent chloride probe.Ambroxol significantly stimulated Cl− efflux from CFBE cells (a sixfold increase after 8 h treatment), and enhanced theexpression of the mRNA of CFTR and a-ENaC, and of the CFTR protein. No significant difference was observed in b-ENaCafter exposure to ambroxol, whereasmRNA expression of g-ENaC was reduced. No significant effects of ambroxol on the ENaCsubunits were observed by Western blot. Ambroxol did not significantly affect the intracellular Ca2+ concentration.Upregulation of CFTR and enhanced Cl efflux after ambroxol treatment should promote transepithelial ion and watertransport, which may improve hydration of the mucus, and therefore be beneficial to CF-patients.

Place, publisher, year, edition, pages
Hoboken, USA: Wiley-Blackwell, 2013
Keywords
Airway epithelium, ambroxol, cystic fibrosis, Cl− efflux, CFTR; ENaC
National Category
Medical and Health Sciences Cell Biology
Research subject
Biomedicine
Identifiers
urn:nbn:se:oru:diva-32769 (URN)10.1002/cbin.10146 (DOI)000325489500002 ()23765701 (PubMedID)2-s2.0-84885861654 (Scopus ID)
External cooperation:
Funder
Swedish Heart Lung FoundationSwedish Research Council
Note

Funding Agencies:

Swedish Science Research Council 

Available from: 2013-12-13 Created: 2013-12-13 Last updated: 2017-12-06Bibliographically approved
Oliynyk, I., Hussain, R., Amin, A., Johannesson, M. & Roomans, G. M. (2013). The effect of NO-donors on chloride efflux, intracellular Ca2+ concentration and mRNA expression of CFTR and ENaC in cystic fibrosis airway epithelial cells. Experimental and molecular pathology (Print), 94(3), 474-480
Open this publication in new window or tab >>The effect of NO-donors on chloride efflux, intracellular Ca2+ concentration and mRNA expression of CFTR and ENaC in cystic fibrosis airway epithelial cells
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2013 (English)In: Experimental and molecular pathology (Print), ISSN 0014-4800, E-ISSN 1096-0945, Vol. 94, no 3, p. 474-480Article in journal (Refereed) Published
Abstract [en]

Since previous studies showed that the endogenous bronchodilator, S nitrosglutathione (GSNO), caused amarked increase in CFTR-mediated chloride (Cl−) efflux and improved the trafficking of CFTR to the plasmamembrane, and that also the nitric oxide (NO)-donor GEA3162 had a similar, but smaller, effect on Cl− efflux, itwas investigatedwhether the NO-donor properties of GSNOwere relevant for its effect on Cl− efflux fromairwayepithelial cells. Hence, the effect of a number of other NO-donors, sodium nitroprusside (SNP), S-nitroso-Nacetyl-DL-penicillamine (SNAP), diethylenetriamine/nitric oxide adduct (DETA-NO), and diethylenetriamine/nitricoxide adduct (DEA-NONOate) on Cl− efflux from CFBE (ΔF508/ΔF508-CFTR) airway epithelial cells was tested.Cl− efflux was determined using the fluorescent N-(ethoxycarbonylmethyl)-6-methoxyquinoliniu bromide(MQAE)-technique. Possible changes in the intracellular Ca2+ concentration were tested by the fluorescent fluo-4method in a confocal microscope system. Like previously with GSNO, after 4 h incubation with the NO-donor, anincreased Cl− efflux was found (in the order SNAP > DETA-NO > SNP). The effect of DEA-NONOate on Cl− effluxwas not significant, and the compound may have (unspecific) deleterious effects on the cells. Again, as withGSNO, after a short (5 min) incubation, SNP had no significant effect on Cl− efflux. None of the NO-donors thathad a significant effect on Cl− efflux caused significant changes in the intracellular Ca2+ concentration. After 4 hpreincubation, SNP caused a significant increase in the mRNA expression of CFTR. SNAP and DEA-NONOatedecreased the mRNA expression of all ENaC subunits significantly. DETA-NO caused a significant decrease only inα-ENaC expression. After a short preincubation, none of the NO-donors had a significant effect, neither on theexpression of CFTR, nor on that of the ENaC subunits in the presence and absence of L-cysteine. It can be concludedthat the effect of GSNO on Cl−efflux is, at least in part, due to its properties as an NO-donor, and the effect is likely tobe mediated by CFTR, not by Ca2+-activated Cl− channels.

Place, publisher, year, edition, pages
Maryland Heights, USA: Elsevier, 2013
Keywords
Nitric oxide donors, Cystic fibrosis, Airway epithelium, CFTR, ENaC, Calcium
National Category
Medical and Health Sciences Cell Biology
Research subject
Biomedicine
Identifiers
urn:nbn:se:oru:diva-32767 (URN)10.1016/j.yexmp.2013.03.003 (DOI)000319535900008 ()23523754 (PubMedID)2-s2.0-84876344605 (Scopus ID)
Funder
Swedish Heart Lung FoundationNIH (National Institute of Health)Swedish Research Council
Available from: 2013-12-13 Created: 2013-12-13 Last updated: 2017-12-06Bibliographically approved
Servetnyk, Z., Jiang, S., Hjelte, L., Gaston, B., Roomans, G. M. & Dragomir, A. (2011). The effect of S-nitrosoglutathione and L-cysteine on chloride efflux from cystic fibrosis airway epithelial cells. Experimental and molecular pathology (Print), 90(1), 79-83
Open this publication in new window or tab >>The effect of S-nitrosoglutathione and L-cysteine on chloride efflux from cystic fibrosis airway epithelial cells
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2011 (English)In: Experimental and molecular pathology (Print), ISSN 0014-4800, E-ISSN 1096-0945, Vol. 90, no 1, p. 79-83Article in journal (Refereed) Published
Abstract [en]

The endogenous bronchodilator, S-nitrosoglutathione (GSNO), has been proposed as a possible pharmacological remedy that reverses the Delta F508-CFTR (cystic fibrosis transmembrane conductance regulator) maturation defect and increases CFTR-mediated chloride efflux in cultured cystic fibrosis airway epithelial cells (CFBE41o(-)). It has also been reported that L-cysteine enhanced S-nitrosothiol uptake and increased the intracellular S-nitrosothiol levels, likely through transnitrosation chemistry. The present study investigated whether L-cysteine augmented the effect of GSNO on chloride efflux from CF airway epithelial cells. Treatment with 10 mu M GSNO combined with 20 mu M L-cysteine resulted in increased chloride efflux from CFBE41o(-) cells after 5 minutes exposure compared to the control efflux rate and to the efflux rate in the presence of L-cysteine alone as measured using the fluorescent dye N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE). Chloride efflux rates from these cells after 4 h exposure to GSNO and L-cysteine were not different from control. Treatment with 10 mu M GSNO alone increased chloride efflux from CFBE41o(-) cells after 4 h but not at shorter incubation times. GSNO with or without L-cysteine did not alter epithelial tight junction integrity. In conclusion, a combination of GSNO with L-cysteine led to significant increase in chloride efflux in CFBE41o(-) cells but the effect was transient and not sustained beyond minutes. (C) 2010 Elsevier Inc. All rights reserved.

National Category
Clinical Medicine
Research subject
Biomedicine
Identifiers
urn:nbn:se:oru:diva-18793 (URN)10.1016/j.yexmp.2010.10.005 (DOI)000286349300013 ()
Available from: 2011-09-29 Created: 2011-09-29 Last updated: 2017-12-08Bibliographically approved
Nilsson, H. E., Dragomir, A., Lazorova, L., Johannesson, M. & Roomans, G. M. (2010). CFTR and tight junctions in cultured bronchial epithelial cells. Experimental and molecular pathology (Print), 88(1), 118-127
Open this publication in new window or tab >>CFTR and tight junctions in cultured bronchial epithelial cells
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2010 (English)In: Experimental and molecular pathology (Print), ISSN 0014-4800, E-ISSN 1096-0945, Vol. 88, no 1, p. 118-127Article in journal (Refereed) Published
Abstract [en]

Airway epithelial salt and water transport takes place through paracellular and transcellular pathways. This transport depends critically on the epithelial sodium channel (ENaC) and the cystic fibrosis transmembrane conductance regulator (CFTR), operating in concert with the paracellular pathway through the tight junctions (TJ). Normal (16HBE14o-), cystic fibrosis (CFBE41o-), and corrected CFBE41o- (CFBE41o-pCep4 overexpressing wtCFTR) airway epithelial cell lines were cultured under isotonic conditions. Transepithelial electrical resistance (TEER) was measured as indicator of the tightness of the cultures. Morphology was investigated by immunofluorescence and paracellular permeability by lanthanum nitrate or [C-14] mannitol as permeability markers. The CFTR-defective cell line CFBE41o- developed higher TEER than its corrected counterpart CFBE41o-pCep4. Addition of a specific inhibitor of CFTR (CFTRinh-172) to 16HBE14o- and CFBE41o-pCep4 cells resulted in a time-dependent increase in TEER, whereas stimulation of CFTR by IBMX and forskolin caused a decrease. Permeability to lanthanum and [C-14] mannitol was lower in CFBE41o- and in 16HBE14o- cells exposed to CFTRinh-172, compared to untreated 16HBE14o- and CFBE41o-pCep4 cells, respectively. 16HBE14o- cells exposed to IBMX and forskolin showed higher permeability to lanthanum but lower permeability to [C-14] mannitol compared to control. Immunofluorescence revealed a disorganization of F-actin and alpha-tubulin in 16HBE14o- cells and CFBE41o- pCep4 exposed to CFTRinh-172 and in CFBE41o- cells. Changes in F-actin and alpha-tubulin in 16HBE14o- cells exposed to IBMX and forskolin were also seen. These results suggest the possibility of an interaction between CFTR and the TJ protein complex, probably via the cytoskeleton. (C) 2009 Elsevier Inc. All rights reserved.

Keywords
CFTR, Transepithelial electrical resistance, Tight junctions, Cytoskeleton
National Category
Medical and Health Sciences
Research subject
Medicine
Identifiers
urn:nbn:se:oru:diva-13020 (URN)10.1016/j.yexmp.2009.09.018 (DOI)000274500100017 ()
Available from: 2011-01-03 Created: 2011-01-03 Last updated: 2017-12-11Bibliographically approved
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